Embedding vitrification ultralow-temperature storage method for bletilla striata seeds

A technology of vitrification ultra-low temperature and preservation method, applied in the field of ultra-low temperature preservation of plant germplasm resources, to achieve high germination rate, shorten dehydration time, and achieve long-term ultra-low temperature preservation

Active Publication Date: 2014-09-03
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the cryopreservati

Method used

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  • Embedding vitrification ultralow-temperature storage method for bletilla striata seeds

Examples

Experimental program
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Example Embodiment

[0031] Example 1

[0032] Collect whole and uncracked white and mature fruits more than 150 days after pollination, rinse them with tap water first, wipe the surface with 75% ethanol, place them on the ultra-clean workbench, and then soak in 75% ethanol for 30s, rinse with sterile water for 2-3 Seconds, followed by soaking with 0.1% mercury liters for 20 minutes, rinsed with sterile water for 4-5 times. Place the sterilized fruit in a sterile petri dish, absorb the surface moisture with sterile filter paper, split the pod longitudinally, and take the powdered seeds in the pod for use.

[0033] Put the powdered seeds into the sodium alginate embedding solution, stir well with a glass rod to evenly distribute the seeds in it. The formula of the sodium alginate embedding solution is: VW+2% sodium alginate+2.0mol·L -1 Glycerin+0.4mol·L -1 Sucrose; use a dropper to drop the sodium alginate embedding solution containing seeds into the calcium ion embedding solution, the formula of the ca...

Example Embodiment

[0038] Example 2-8

[0039] In the implementation steps of Examples 2-8, the dehydration time in the PVS2 solution was 40 min, 60 min, 80 min, 100 min, 120 min, 140 min, 160 min, and the other steps were the same as in Example 1.

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Abstract

The invention relates to an embedding vitrification ultralow-temperature storage method for bletilla striata seeds. The method comprises the following steps: collecting integral fruit, cleaning, wiping surfaces with ethanol, immersing in ethanol, flushing with sterile water, immersing in a mercuric chloride solution with a concentration of 0.1%, flushing, absorbing water on the fruit surfaces, splitting capsules in a longitudinal direction, and taking out powder-like seeds; pouring seeds into a sodium alginate embedding solution, stirring uniformly, dropwise putting into a calcium ion embedding solution, and putting in a room-temperature environment and forming embedded beads through solidifying; putting the embedded beads into a PVS2 solution and performing dewatering processing for 20-160 min; putting into a refrigeration tube, adding a PVS2 solution, and then putting the refrigeration tube in liquid nitrogen for refrigeration storage; when seeds are needed, taking out the refrigeration tube, unfreezing and removing the PVS2 solution, and washing; and inoculating the washed embedded beads to a solid VW medium for culture, enabling the seed to germinate, and performing domestication transplanting after seedlings grow to have the height of 6-10 cm. The seeds obtained by employing the method are high in germination rate and the cost can be saved.

Description

technical field [0001] The invention relates to a method for cryopreservation of plant germplasm resources, in particular to a method for embedding and vitrification cryopreservation of seeds of bletilla germplasm. Background technique [0002] Baiji (Bletilla striata (Thunb.) Reichb.f.), also known as Lianjicao, Ruolan, Gangen, Baigen, Baili, Baiji, and Zihui, is a perennial herbaceous plant belonging to Orchidaceae and is distributed in Asia. Northern Myanmar passes through China to Japan and the Korean Peninsula. Baiji has purple inflorescences and dense leaves. It can be cultivated as a potted ornamental plant or as a ground cover. It can also produce fresh cut flowers and has good ornamental value. The tuber of Baiji is a commonly used traditional Chinese medicine, which has the functions of astringent and hemostasis, detumescence and granulation promotion, and promotion of wound healing. Significant curative effect. In recent years, the scope of clinical application...

Claims

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Application Information

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IPC IPC(8): A01N3/00
Inventor 王彩霞田敏
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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